The ABC transporter MsbA adopts the wide inward-open conformation in E. coli cells
Autor(en): | Galazzo, Laura Meier, Gianmarco Januliene, Dovile Parey, Kristian De Vecchis, Dario Striednig, Bianca Hilbi, Hubert Schaefer, V, Lars Kuprov, Ilya Moeller, Arne Bordignon, Enrica Seeger, Markus A. |
Stichwörter: | ATP-BINDING; BINDING CASSETTE TRANSPORTER; CRYO-EM; CRYSTAL-STRUCTURE; ESCHERICHIA-COLI; FLEXIBILITY; MOLECULAR-DYNAMICS; Multidisciplinary Sciences; NANOBODIES; P-GLYCOPROTEIN REVEAL; Science & Technology - Other Topics; STRUCTURAL BASIS | Erscheinungsdatum: | 2022 | Herausgeber: | AMER ASSOC ADVANCEMENT SCIENCE | Journal: | SCIENCE ADVANCES | Volumen: | 8 | Ausgabe: | 41 | Zusammenfassung: | Membrane proteins are currently investigated after detergent extraction from native cellular membranes and reconstitution into artificial liposomes or nanodiscs, thereby removing them from their physiological environment. However, to truly understand the biophysical properties of membrane proteins in a physiological environment, they must be investigated within living cells. Here, we used a spin-labeled nanobody to interrogate the conformational cycle of the ABC transporter MsbA by double electron-electron resonance. Unexpectedly, the wide inward-open conformation of MsbA, commonly considered a nonphysiological state, was found to be prominently populated in Escherichia coli cells. Molecular dynamics simulations revealed that extensive lateral portal opening is essential to provide access of its large natural substrate core lipid A to the binding cavity. Our work paves the way to investigate the conformational landscape of membrane proteins in cells. |
ISSN: | 2375-2548 | DOI: | 10.1126/sciadv.abn6845 |
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geprüft am 13.05.2024